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Analysis of CBL Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0046-LGZ)

Introduction

Official Full Name: Cbl proto-oncogene
Also known as: CBL2; NSLL; C-CBL; RNF55; FRA11B
This gene is a proto-oncogene encoding RING finger E3 ubiquitin ligase. The encoded protein is one of the enzymes required by the proteasome to degrade substrates. This protein mediates the transfer of ubiquitin from ubiquitin-conjugating enzymes (E2) to specific substrates. The protein also contains an N-terminal phosphotyrosine-binding domain that enables it to interact with many tyrosine-phosphorylated substrates and target them for proteasomal degradation. Therefore, it acts as a negative regulator in many signal transduction pathways. This gene has been found to be mutated or translocated in many cancers including acute myeloid leukaemia, and expansion of CGG repeats in the 5' UTR has been associated with Jacobsen syndrome. Mutations in this gene are also the cause of Noonan syndrome-like dis order.




Principle

Under certain conditions, two single strands of nucleic acid with certain homology can be specifically hybridized to form double strands according to the principle of base complementarity. Generally, DNA molecules to be detected are digested with restriction enzymes, separated by agar-gel electrophoresis, denatured and transferred to nitrocellulocellulose film or nylon film or other solid phase support according to their position in the gel, fixed and then reacted with DNA probes labeled with isotopes or other markers. This is followed by autoradiography or an enzyme reaction to detect the amount of specific DNA molecules. If the object to be tested contains a sequence that is complementary to the probe, the two are combined by the principle of base complementarity, and the free probe is washed and detected by self-development or other suitable techniques, thus revealing the fragment to be tested and its relative size.

Applications

Gene Rearrangement Detection

Procedure

1. Sample Processing
2. DNA Extraction and Digestion
3. Gel Electrophoresis
4. Gel Pretreatment
5. Transfer membrane
6. Probe Labeling
7. Prehybridization (blocking)
8. Southern hybridization
9. Membrane washing
10. Autoradiographic Assay
11. Results Analysis

Materials

Sample: DNA, Bacterial Fluid/Tissue/Cell
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